React and act: a qualitative study of how nursing home leaders follow up on staff-to-resident abuse.
Elder abuse in nursing homes is a complex multifactorial problem and entails various associations across personal, social, and organisational factors. One way leaders can prevent abuse and promote quality and safety for residents is to follow up on any problems that may arise in clinical practice in a way that facilitates learning. How nursing home leaders follow up and what they follow up on might reflect their perceptions of abuse, its causal factors, and the prevention strategies used in the nursing home. The aim of this study was to explore how nursing home leaders follow up on reports and information regarding staff-to-resident abuse. A qualitative explorative design was used. The sample comprised 43 participants from two levels of nursing home leadership representing six municipalities and 21 nursing homes in Norway. Focus group interviews were conducted with 28 care managers, and individual interviews took place with 15 nursing home directors. The constant comparative method was used for the analyses. Nursing home leaders followed up incidents of staff-to-resident abuse on three different levels as follows: 1) on an individual level, leaders performed investigations and meetings, guidance, supervision, and occasionally relocated staff members; 2) on a group level, feedback, openness, and reflection for shared understanding were strategies leaders used; and 3) on an organisational level, the main solutions were to adjust to available resources, training, and education. We found that leaders had difficulties defining harm and a perceived lack of power to follow up on all levels. In addition, they did not have adequate tools for evaluating the effect of the measures that were taken. Nursing home leaders need to be clear about how they should follow up incidents of elder abuse on different levels in the organisation and about their role in preventing elder abuse. Evaluation tools that facilitate systematic organisational learning are needed. Nursing homes must operate as open, blame-free cultures that acknowledge that incidents of elder abuse in patient care arise not only from the actions of individuals but also from the complex everyday life of which they are a part and in which they operate.


Patterns of Care and Outcomes of Elderly Esophageal Cancer Patients Not Meeting Age-based Criteria of the CROSS Trial.
The CROSS trial established neoadjuvant chemoradiation followed by surgery (nCRT-S) as superior to surgery alone (S) for locally advanced esophageal cancer (EC). However, because patients above 75 years of age were excluded, this comparison cannot be extrapolated to older patients. This study of a large, contemporary national database evaluated practice patterns in elderly patients ineligible for CROSS, and analyzed overall survival (OS) between nCRT+S, S, and definitive CRT (dCRT). The National Cancer Data Base was queried for EC patients with cT1N1M0/T2-3N0-1M0 EC (per the CROSS trial) but 76 years and above of age. Multivariable logistic regression ascertained factors associated with nCRT+S (vs. S). Kaplan-Meier analysis evaluated OS; Cox multivariate analysis determined variables associated with OS. Propensity matching aimed to address group imbalances and indication biases. Of 4099 total patients, 594 (14%) underwent nCRT+S, 494 (12%) underwent S, and 3011 (73%) underwent dCRT. Since 2010, trimodality management has risen, corresponding to declines in S and dCRT. Median OS in the respective groups were 26.7, 20.3, and 17.8 months (P<0.05). Following propensity matching, there was a trend towards higher OS with nCRT-S over S (P=0.077); dCRT showed poorer OS than nCRT-S (P<0.001) but was equivalent to S (P=0.669). Before and following matching, nCRT-S experienced equivalent 30- and 90-day mortality as S (P>0.05), with lower 30-day readmission and postoperative hospital stay (P<0.05). Although most older patients not meeting CROSS criteria undergo dCRT, utilization of trimodality therapy is rising. Despite the trend towards higher OS with trimodality therapy without poorer postoperative outcomes, careful patient selection continues to be essential in this population.


Protective role of the Mycobacterium smegmatis IdeR against reactive oxygen species and isoniazid toxicity.
To understand the mechanism by which IdeR is necessary for maintaining wild type levels of KatG and SodA enzyme activity and normal isoniazid (INH) resistance. To identify the step(s) of SodA and KatG function that were affected by the ideR mutation, quantitative western immunoassays and ribonucleic acid (RNA) hybridizations were performed. To see if the increased INH sensitivity of the ideR mutant was caused by lower SodA activity, the Mycobacterium smegmatis sod gene was inactivated. The levels of KatG and SodA mRNA and protein in the M. smegmatis IdeR mutant are decreased to approximately 20-40% of those observed in the wild type parent strain. This is quantitatively similar to the decrease in KatG and SodA enzyme activities originally observed in the ideR strain. The M. smegmatis sodA mutant was slightly more sensitive to INH, compared to the wild type strain and was more resistant than the ideR mutant. IdeR is necessary for full expression of the M. smegmatis katG and sodA genes. It is not yet known whether this protein acts directly at the gene level. The lower levels of SodA contribute slightly to the increased susceptibility to INH of the ideR mutant, but cannot explain the magnitude of the INH sensitivity observed when IdeR is not present. These data suggest that IdeR is a regulator of the cellular stress response, as it has a protective role in cells facing environmental stresses, such as increased levels of reactive oxygen species and INH toxic intermediates. These conclusions do not necessarily apply to IdeR's role in M. tuberculosis physiology, since we have not inactivated its gene in this pathogen.


Fli1 acts at the top of the transcriptional network driving blood and endothelial development.
Blood and endothelium arise in close association during development, possibly from a common precursor, the hemangioblast [1-4]. Genes essential for blood and endothelial development contain functional ETS binding sites, and binding and expression data implicate the transcription factor, friend leukaemia integration 1 (Fli1) [5-10]. However, loss-of-function phenotypes in mice, although suffering both blood and endothelial defects, have thus far precluded the conclusion that Fli1 is essential for these two lineages [11, 12]. By using Xenopus and zebrafish embryos, we show that loss of Fli1 function results in a substantial reduction or absence of hemangioblasts, revealing an absolute requirement. TUNEL assays show that the cells are eventually lost by apoptosis, but only after the regulatory circuit has been disrupted by loss of Fli1. In addition, a constitutively active form of Fli1 is sufficient to induce expression of key hemangioblast genes, such as Scl/Tal1, Lmo2, Gata2, Etsrp, and Flk1. Epistasis assays show that Fli1 expression is induced by Bmp signaling or Cloche, depending on the hemangioblast population, and in both cases Fli1 acts upstream of Gata2, Scl, Lmo2, and Etsrp. Taken together, these results place Fli1 at the top of the transcriptional regulatory hierarchy for hemangioblast specification in vertebrate embryos.


Characterization and drug release investigation of amorphous drug-hydroxypropyl methylcellulose composites made via supercritical carbon dioxide assisted impregnation.
Hydroxypropylmethyl cellulose (HPMC)-indomethacin (4:1, w/w) drug composites (DCs) were prepared via supercritical carbon dioxide (sc-CO(2)) assisted impregnation. The effect of processing temperature (at fixed pressures) on the physical and other properties of the resulting HPMC-indomethacin DCs was investigated using a range of analytical techniques, including differential scanning calorimetry (DSC), Fourier transform infrared (FTIR) spectroscopy, Raman spectroscopy, and powder X-ray diffraction (XRD) methods. The data suggest that for a 4:1 (w/w) HPMC-indomethacin ratio prepared at 130 degrees C (17.2 MPa), the indomethacin exists entirely in an amorphous dispersion within the polymer matrix. The primary interaction between HPMC and indomethacin appears to be hydrogen bonding between the carboxylic acid carbonyl group of indomethacin and hydroxyl group of HPMC. The initial (first 15 min) and overall drug release behavior within a 5h timeframe for the HPMC-indomethacin DCs, was analyzed. For the HPMC-indomethacin drug composite processed at 130 degrees C/17.2 MPa, drug release behavior obeyed a n-power law (n=0.54).


REST represses a subset of the pancreatic endocrine differentiation program.
To contribute to devise successful beta-cell differentiation strategies for the cure of Type 1 diabetes we sought to uncover barriers that restrict endocrine fate acquisition by studying the role of the transcriptional repressor REST in the developing pancreas. Rest expression is prevented in neurons and in endocrine cells, which is necessary for their normal function. During development, REST represses a subset of genes in the neuronal differentiation program and Rest is down-regulated as neurons differentiate. Here, we investigate the role of REST in the differentiation of pancreatic endocrine cells, which are molecularly close to neurons. We show that Rest is widely expressed in pancreas progenitors and that it is down-regulated in differentiated endocrine cells. Sustained expression of REST in Pdx1(+) progenitors impairs the differentiation of endocrine-committed Neurog3(+) progenitors, decreases beta and alpha cell mass by E18.5, and triggers diabetes in adulthood. Conditional inactivation of Rest in Pdx1(+) progenitors is not sufficient to trigger endocrine differentiation but up-regulates a subset of differentiation genes. Our results show that the transcriptional repressor REST is active in pancreas progenitors where it gates the activation of part of the beta cell differentiation program.


Cerebrospinal fluid matrix metalloproteinases are elevated in cerebral adrenoleukodystrophy and correlate with MRI severity and neurologic dysfunction.
X-linked adrenoleukodystrophy results from mutations in the ABCD1 gene disrupting the metabolism of very-long-chain fatty acids. The most serious form of ALD, cerebral adrenoleukodystrophy (cALD), causes neuroinflammation and demyelination. Neuroimaging in cALD shows inflammatory changes and indicates blood-brain-barrier (BBB) disruption. We hypothesize that disruption may occur through the degradation of the extracellular matrix defining the BBB by matrix metalloproteinases (MMPs). MMPs have not been evaluated in the setting of cALD. We used a multiplex assay to correlate the concentration of MMPs in cerebrospinal fluid and plasma to the severity of brain inflammation as determined by the ALD MRI (Loes) score and the neurologic function score. There were significant elevations of MMP2, MMP9, MMP10, TIMP1, and total protein in the CSF of boys with cALD compared to controls. Levels of MMP10, TIMP1, and total protein in CSF showed significant correlation [p<0.05 for each with pre-transplant MRI Loes Loes scores (R(2) = 0.34, 0.20, 0.55 respectively). Levels of TIMP1 and total protein in CSF significantly correlated with pre-transplant neurologic functional scores (R(2) = 0.22 and 0.48 respectively), and levels of MMP10 and total protein in CSF significantly correlated with one-year post-transplant functional scores (R(2) = 0.38 and 0.69). There was a significant elevation of MMP9 levels in plasma compared to control, but did not correlate with the MRI or neurologic function scores. MMPs were found to be elevated in the CSF of boys with cALD and may mechanistically contribute to the breakdown of the blood-brain-barrier. MMP concentrations directly correlate to radiographic and clinical neurologic severity. Interestingly, increased total protein levels showed superior correlation to MRI score and neurologic function score before and at one year after transplant.


Is ceramide signaling a target for vascular therapeutic intervention?
Today's society is plagued by cardiovascular diseases such as hypertension and atherosclerosis. Research devoted to the study of these diseases has focused, in part, on physiological phenomena responsible for the development and progression of the disease. Peripheral vascular function is one such focal point of research, and identification of cellular mechanisms that regulate vascular contractility and/or cellular proliferation is crucial for the development of new therapeutic interventions to combat these diseases. This review evaluates a new signaling mechanism, the ceramide signaling pathway as a potential target for therapeutic intervention. Special attention is given ceramide signaling in the contexts of vascular reactivity and cell proliferation in the vasculature. While ceramide signaling is a nascent area of vascular research, a growing body of evidence from other physiological systems implicates this new pathway as a potential regulator of contractile and cell proliferative functions within the blood vessel wall.


Berberine protects the liver and kidney against functional disorders and histological damages induced by ferrous sulfate.
Iron is an essential element for living organisms. Iron overload can have detrimental effects on health. This study pertains to the protective role of berberine against ferrous sulfate-induced hepatic and renal functional disorders and histological damages in rats. The rats were divided into four groups (n=7): Sham, Ber (10 mg/kg/day for 14 days, by gavage), FS (ferrous sulfate, 30 mg/kg/day for 14 days, intraperitoneally), FS + Ber (ferrous sulfate, 30 mg/kg/day for 14 days; berberine, 10 mg/kg/day for 11 days from fourth day of ferrous sulfate injection). After 24 hr, blood, urine, and tissue samples were collected. Compared with sham and Ber groups, administration of ferrous sulfate resulted in liver and kidney dysfunction as evidenced by significantly higher levels of serum hepatic markers and bilirubin, and lower levels of serum albumin, total protein, triglyceride, cholesterol, and glucose, as well as lower creatinine clearance and higher fractional excretion of sodium. This was accompanied by increased malondialdehyde levels and histological damages. Berberine treatment significantly reversed the levels of serum hepatic markers, renal functional markers and lipid peroxidation marker in the FS + Ber group. Furthermore, it restored the levels of serum total protein, albumin, glucose, triglycerides, and cholesterol with a decrease in bilirubin concentration in the blood. All these changes were corroborated by histological observations of the liver and kidney. Berberine protects the liver and kidneys against ferrous sulfate-induced toxicity by reduction in lipid peroxidation and ability to chelate iron.


Prevalence of transportation safety measures portrayed in primetime US television programs and commercials.
To determine the prevalence of transportation related safety behaviors, such as seatbelt and helmet use, in primetime US television programs and commercials. Cross sectional study. Top rated television programs and associated commercials from four major US television networks were reviewed for the prevalence of transportation safety related behaviors during a one month period in 2005. Programs were categorized according to the time and network of airing, program type, program rating, and--for commercials--type of product being advertised Occupants of automobiles, motorcycles, or bicycles in 507 instances in which a transportation scene was aired. Seatbelt use was depicted in 62% and 86% of individuals in television program and commercial automobile scenes, respectively. The prevalence of motorcycle helmet use was 47% in television programs and 100% in commercials. Bicycle helmets were used in 9% of television programs and 84% of commercials. The frequency of seatbelt use in programs and commercials varied by television rating and genre but did not differ by network, time of airing, or age of character portrayed. The prevalence of safety related behaviors aired on major US networks during primetime slots is higher than previous reports but still much lower than national averages. Commercials, in contrast, portray transportation safety measures with a frequency that exceeds that of US television programs or most national surveys.


Two patients with eating disorders treated by naltrexone.
According to a recent literature review on the opioid mechanism in eating disorders, we found that there is increasing reason to re-examine the treatment potential of naltrexone. The endogenous opioid system belongs to the important modulators of food intake. The eating disorders share many traits with substance dependence models. We present two case histories of time-limited naltrexone therapy to show that, in clinical practice, individualized indication may contribute to short-term improvement and to prediction of a different long-term treatment outcome.


Sequences of two related multiple antibiotic resistance virulence plasmids sharing a unique IS26-related molecular signature isolated from different Escherichia coli pathotypes from different hosts.
Enterohemorrhagic Escherichia coli (EHEC) and atypical enteropathogenic E. coli (aEPEC) are important zoonotic pathogens that increasingly are becoming resistant to multiple antibiotics. Here we describe two plasmids, pO26-CRL125 (125 kb) from a human O26:H- EHEC, and pO111-CRL115 (115kb) from a bovine O111 aEPEC, that impart resistance to ampicillin, kanamycin, neomycin, streptomycin, sulfathiazole, trimethoprim and tetracycline and both contain atypical class 1 integrons with an identical IS26-mediated deletion in their 3´-conserved segment. Complete sequence analysis showed that pO26-CRL125 and pO111-CRL115 are essentially identical except for a 9.7 kb fragment, present in the backbone of pO26-CRL125 but absent in pO111-CRL115, and several indels. The 9.7 kb fragment encodes IncI-associated genes involved in plasmid stability during conjugation, a putative transposase gene and three imperfect repeats. Contiguous sequence identical to regions within these pO26-CRL125 imperfect repeats was identified in pO111-CRL115 precisely where the 9.7 kb fragment is missing, suggesting it may be mobile. Sequences shared between the plasmids include a complete IncZ replicon, a unique toxin/antitoxin system, IncI stability and maintenance genes, a novel putative serine protease autotransporter, and an IncI1 transfer system including a unique shufflon. Both plasmids carry a derivate Tn21 transposon with an atypical class 1 integron comprising a dfrA5 gene cassette encoding resistance to trimethoprim, and 24 bp of the 3´-conserved segment followed by Tn6026, which encodes resistance to ampicillin, kanymycin, neomycin, streptomycin and sulfathiazole. The Tn21-derivative transposon is linked to a truncated Tn1721, encoding resistance to tetracycline, via a region containing the IncP-1α oriV. Absence of the 5 bp direct repeats flanking Tn3-family transposons, indicates that homologous recombination events played a key role in the formation of this complex antibiotic resistance gene locus. Comparative sequence analysis of these closely related plasmids reveals aspects of plasmid evolution in pathogenic E. coli from different hosts.


Vestibular Migraine: How to Sort it Out and What to Do About it.
Vestibular migraine (VM) is the most common neurologic cause of vertigo in adults and results in significant utilization of health care resources, but remains under-recognized and underdiagnosed. Review of literature in PubMed using the following terms: vestibular migraine, migraine-associated vertigo, vertiginous migraine, benign recurrent vertigo, migraine-associated dizziness, migraine, migraine treatment, Meniere disease (MD), vertebrobasilar ischemia (VBI), posterior circulation stroke, benign paroxysmal positional vertigo, and episodic-ataxia Type 2 (EA2). VM can manifest with a variety of vestibular symptoms, including spontaneous vertigo, triggered vertigo, positional vertigo, and head-motion dizziness. Patients may report more than 1 vestibular symptom. Episodes of vertigo are often, but not always, accompanied by headache. Auditory symptoms are frequently associated with VM attacks and may mimic the manifestations of MD. Other migrainous features that accompany VM attacks include photophobia, phonophobia, osmophobia, and visual aura. Interictally, patients may suffer from persistent dizziness or isolated paroxysmal vestibular symptoms. Mood disorders (particularly anxiety) are often found in VM. Abnormal neuro-otologic findings are not uncommon in patients with VM. Differential diagnoses for VM include MD, VBI, EA2, and migraine with brainstem aura. For rescue treatment, triptans, vestibular suppressants, and/or antiemetic agents may be considered. Pharmacologic migraine preventives (antiepileptics, beta-blockers, and antidepressants) are often useful. The keys to correctly diagnosing VM is identifying a relationship between vestibular symptoms and migrainous features and being aware of the heterogeneity of manifestations of this enigmatic, but treatable, condition. The principles of treatment of VM include rescue therapy, lifestyle modification, nonpharmacologic migraine preventives, pharmacologic migraine prophylaxis, and treatment of comorbidities.


Antibody-Mediated Protective Mechanisms Induced by a Trivalent Parainfluenza Virus-Vectored Ebolavirus Vaccine.
Ebolaviruses Zaire (EBOV), Bundibugyo (BDBV), and Sudan (SUDV) cause human disease with high case fatality rates. Experimental monovalent vaccines, which all utilize the sole envelope glycoprotein (GP), do not protect against heterologous ebolaviruses. Human parainfluenza virus type 3-vectored vaccines offer benefits, including needle-free administration and induction of mucosal responses in the respiratory tract. Multiple approaches were taken to induce broad protection against the three ebolaviruses. While GP consensus-based antigens failed to elicit neutralizing antibodies, polyvalent vaccine immunization induced neutralizing responses to all three ebolaviruses and protected animals from death and disease caused by EBOV, SUDV, and BDBV. As immunization with a cocktail of antigenically related antigens can skew the responses and change the epitope hierarchy, we performed comparative analysis of antibody repertoire and Fc-mediated protective mechanisms in animals immunized with monovalent versus polyvalent vaccines. Compared to sera from guinea pigs receiving the monovalent vaccines, sera from guinea pigs receiving the trivalent vaccine bound and neutralized EBOV and SUDV at equivalent levels and BDBV at only a slightly reduced level. Peptide microarrays revealed a preponderance of binding to amino acids 389 to 403, 397 to 415, and 477 to 493, representing three linear epitopes in the mucin-like domain known to induce a protective antibody response. Competition binding assays with monoclonal antibodies isolated from human ebolavirus infection survivors demonstrated that the immune sera block the binding of antibodies specific for the GP glycan cap, the GP1-GP2 interface, the mucin-like domain, and the membrane-proximal external region. Thus, administration of a cocktail of three ebolavirus vaccines induces a desirable broad antibody response, without skewing of the response toward preferential recognition of a single virus.IMPORTANCE The symptoms of the disease caused by the ebolaviruses Ebola, Bundibugyo, and Sudan are similar, and their areas of endemicity overlap. However, because of the limited antigenic relatedness of the ebolavirus glycoprotein (GP) used in all candidate vaccines against these viruses, they protect only against homologous and not against heterologous ebolaviruses. Therefore, a broadly specific pan-ebolavirus vaccine is required, and this might be achieved by administration of a cocktail of vaccines. The effects of cocktail administration of ebolavirus vaccines on the antibody repertoire remain unknown. Here, an in-depth analysis of the antibody responses to administration of a cocktail of human parainfluenza virus type 3-vectored vaccines against individual ebolaviruses was performed, which included analysis of binding to GP, neutralization of individual ebolaviruses, epitope specificity, Fc-mediated functions, and protection against the three ebolaviruses. The results demonstrated potent and balanced responses against individual ebolaviruses and no significant reduction of the responses compared to that induced by individual vaccines.


Factors influencing pharmacist performance: a review of the peer-reviewed literature.
To undertake a review of peer-reviewed literature to explore factors affecting pharmacists' performance. The following databases were searched: Medline, Embase, Scopus, ISI Web of Knowledge and PsychInfo. Inclusion criteria were: English language only, published between 1990 and 2010 and published in the United Kingdom (UK), United States of America (USA), Canada, Australia, New Zealand or Europe. The search strategy identified 37 items. The review found that there was some evidence to suggest that pharmacists with certain characteristics (e.g. being male, being of ethnic minority origin, working in community pharmacy and having trained overseas) were more likely to experience performance problems. Factors relating to workload and work environment were associated with performance problems, particularly in relation to errors. There was some limited evidence to suggest that experiencing problems with alcohol or drugs could negatively impact on pharmacists' performance. The findings suggest that pharmacist performance may be affected by multiple factors, including personal characteristics such as age, gender, ethnicity, place of primary qualification, factors associated with the workplace and mental and physical health problems. The evidence is not unequivocal and gaps in the literature exist, suggesting that pharmacist performance is an under-researched area.


The Val/Met polymorphism of the brain-derived neurotrophic factor (BDNF) gene predicts decline in perceptual speed in older adults.
The brain-derived neurotrophic factor (BDNF) promotes activity-dependent synaptic plasticity, and contributes to learning and memory. We investigated whether a common Val66Met missense polymorphism (rs6265) of the BDNF gene is associated with individual differences in cognitive decline (marked by perceptual speed) in old age. A total of 376 participants of the Berlin Aging Study, with a mean age of 83.9 years at first occasion, were assessed longitudinally up to 11 times across more than 13 years on the Digit-Letter task. Met carriers (n = 123, 34%) showed steeper linear decline than Val homozygotes (n = 239, 66%); the corresponding contrast explained 2.20% of the variance in change in the entire sample, and 3.41% after excluding individuals at risk for dementia. These effects were not moderated by sex or socioeconomic status. Results are consistent with the hypothesis that normal aging magnifies the effects of common genetic variation on cognitive functioning.


The role of soil organic carbon in the global cycling of persistent organic pollutants (POPs): interpreting and modelling field data.
Soil is an important global reservoir for persistent organic pollutants (POPs). The interaction between air (which often receives the majority of emissions) and soil plays a key role in the long term environmental cycling and fate of these chemicals. Soil surveys have been carried out to try and estimate regional and global distribution/inventory of POPs. A correlation between soil POPs concentration and soil organic carbon (SOC) has been observed in background soils [Meijer et al., 2003. Global distribution and budget of PCBs and HCB in background surface soils: implications for sources and environmental processes. Environ. Sci. Technol., 37, 667], provoking discussion about whether POPs will approach steady-state (or equilibrium) between air and SOC, on a global scale. This manuscript investigates this relationship and in particular how soil concentrations can be influenced by factors such as temperature, SOC content and physicochemical properties. A simple two box model designed to investigate parameters that are likely to affect air-soil exchange revealed that more volatile chemicals such as HCB are likely to achieve steady-state conditions between air and soil relatively quickly whilst relatively involatile chemicals, such as heavy PCBs, may take considerably longer and other compounds (e.g. OCDD) may never achieve it. These model calculations provide an insight into which fate processes (e.g. volatilisation or degradation) may control a chemicals fate in the terrestrial environment. A different modelling exercise was used to explore the complex interaction of environmental parameters, representative of 'real world' conditions to study their potential influence on POPs cycling at the European scale. Results from the model suggested that compound degradation rates in soil (linked to SOC content), temperature, vegetation cover and ecosystem C turnover are all likely to significantly influence POP air-soil exchange and fate.


Glucocorticoids reduce phobic fear in humans.
Phobias are characterized by excessive fear, cued by the presence or anticipation of a fearful situation. Whereas it is well established that glucocorticoids are released in fearful situations, it is not known whether these hormones, in turn, modulate perceived fear. As extensive evidence indicates that elevated glucocorticoid levels impair the retrieval of emotionally arousing information, they might also inhibit retrieval of fear memory associated with phobia and, thereby, reduce phobic fear. Here, we investigated whether acutely administrated glucocorticoids reduced phobic fear in two double-blind, placebo-controlled studies in 40 subjects with social phobia and 20 subjects with spider phobia. In the social phobia study, cortisone (25 mg) administered orally 1 h before a socio-evaluative stressor significantly reduced self-reported fear during the anticipation, exposure, and recovery phase of the stressor. Moreover, the stress-induced release of cortisol in placebo-treated subjects correlated negatively with fear ratings, suggesting that endogenously released cortisol in the context of a phobic situation buffers fear symptoms. In the spider phobia study, repeated oral administration of cortisol (10 mg), but not placebo, 1 h before exposure to a spider photograph induced a progressive reduction of stimulus-induced fear. This effect was maintained when subjects were exposed to the stimulus again 2 days after the last cortisol administration, suggesting that cortisol may also have facilitated the extinction of phobic fear. Cortisol treatment did not reduce general, phobia-unrelated anxiety. In conclusion, the present findings in two distinct types of phobias indicate that glucocorticoid administration reduces phobic fear.


Ribosomal DNA integrating rAAV-rDNA vectors allow for stable transgene expression.
Although recombinant adeno-associated virus (rAAV) vectors are proving to be efficacious in clinical trials, the episomal character of the delivered transgene restricts their effectiveness to use in quiescent tissues, and may not provide lifelong expression. In contrast, integrating vectors enhance the risk of insertional mutagenesis. In an attempt to overcome both of these limitations, we created new rAAV-rDNA vectors, with an expression cassette flanked by ribosomal DNA (rDNA) sequences capable of homologous recombination into genomic rDNA. We show that after in vivo delivery the rAAV-rDNA vectors integrated into the genomic rDNA locus 8-13 times more frequently than control vectors, providing an estimate that 23-39% of the integrations were specific to the rDNA locus. Moreover, a rAAV-rDNA vector containing a human factor IX (hFIX) expression cassette resulted in sustained therapeutic levels of serum hFIX even after repeated manipulations to induce liver regeneration. Because of the relative safety of integration in the rDNA locus, these vectors expand the usage of rAAV for therapeutics requiring long-term gene transfer into dividing cells.


Low-level and ultralow-volume hollow waveguide based carbon monoxide sensor.
We demonstrate an ultralow sample volume optical carbon monoxide sensor with detection sensitivity of 180 parts in 10(9) (1σ at 1 Hz). The utilization of a 2.3 μm surface-emitting laser directly coupled to a 3 m hollow capillary fiber as the gas cell is proven to be a compact, sensitive, and cost-efficient gas sensing concept. By mechanical vibration of the fiber, an absorbance resolution of 10(-5) is achieved, which is comparable to single-reflective (double-pass) cells. An improvement of sensitivity over the conventional single-reflective cell is thus approximately linearly scaled with the enhancement of the optical path length, which is usually more than 1 order of magnitude.


Treatment of dural fistulas involving the deep cerebral venous system.
Eight patients with dural arteriovenous fistulas involving the deep cerebral venous system were treated by a combination of preoperative embolization, intraoperative embolization, and/or surgical excision. All eight patients were men 30-71 years old (mean age, 48.5). The presenting symptoms were hemorrhage in four patients (two subarachnoid, one intraventricular, and one parenchymal), stroke in two patients, and severe chronic headaches in two patients. Four patients were treated and cured by preoperative embolization of external carotid feeding vessels followed by direct intraoperative placement of liquid adhesives into the fistula site. Two patients underwent preoperative embolization followed by surgical interruption of feeding vessels to the fistula. Both patients had persistent fistulas and were subsequently treated by intraoperative embolization with liquid adhesives. One patient was cured and the second had 95% reduction in fistula size. The remaining two patients had surgical excision of the fistula, one in combination with preoperative embolization. Both were completely cured. Two patients developed hydrocephalus after placement of liquid adhesive into the involved vein of Galen and were successfully treated with placement of ventriculoperitoneal shunts. Follow-up periods ranged from 7 to 21 months (mean, 14). We found that patients with dural arteriovenous fistulas could be treated effectively through a combination of neuroradiologic and surgical intervention.


Parametric blind-deconvolution algorithm to remove image artifacts in hybrid imaging systems.
Hybrid imaging systems employing cubic phase modulation in the pupil-plane enable significantly increased depth of field, but artifacts in the recovered images are a major problem. We present a parametric blind-deconvolution algorithm, based on minimization of the high-frequency content of the restored image that enables recovery of artifact-free images for a wide range of defocus. We show that the algorithm enables robust matching of the image recovery kernel with the optical point-spread function to enable, for the first time, optimally low noise levels in recovered images.


Replication of dengue type 2 virus in Culex quinquefasciatus (Diptera: Culicidae).
We were able to infect Culex quinquefasciatus by the parenteral route with dengue virus type 2. The percentage of mosquitoes infected was dose dependent and we obtained a rate of 45.6% infected Cx. quinquefasciatus when a 10(5.9) MID50 (mosquito infectious dose for 50% of the individuals as measured in Aedes aegypti) of dengue virus type 2 per mosquito was used. Infection was detected by an immunofluorescent assay performed on mosquito head squashes 14 days after infection. The replication of dengue virus in Cx. quinquefasciatus was either at a very low level of magnitude or generated a large number of noninfectious particles since the triturated bodies of infected Cx. quinquefasciatus did not infect Ae. aegypti mosquitoes when inoculated parenterally. We were unable to infect Cx. quinquefasciatus females orally with an artificial meal that infected 100% of Ae. aegypti females. These findings lead us to agree with the consensus that Cx. quinquefasciatus should not be considered a biological vector of dengue viruses.


The stories we tell: qualitative research interviews, talking technologies and the 'normalisation' of life with HIV.
Since the earliest days of the HIV/AIDS epidemic, talking about the virus has been a key way affected communities have challenged the fear and discrimination directed against them and pressed for urgent medical and political attention. Today, HIV/AIDS is one of the most prolifically and intimately documented of all health conditions, with entrenched infrastructures, practices and technologies--what Vinh-Kim Nguyen has dubbed 'confessional technologies'--aimed at encouraging those affected to share their experiences. Among these technologies, we argue, is the semi-structured interview: the principal methodology used in qualitative social science research focused on patient experiences. Taking the performative nature of the research interview as a talking technology seriously has epistemological implications not merely for how we interpret interview data, but also for how we understand the role of research interviews in the enactment of 'life with HIV'. This paper focuses on one crucial aspect of this enactment: the contemporary 'normalisation' of HIV as 'just another' chronic condition--a process taking place at the level of individual subjectivities, social identities, clinical practices and global health policy, and of which social science research is a vital part. Through an analysis of 76 interviews conducted in London (2009-10), we examine tensions in the experiential narratives of individuals living with HIV in which life with the virus is framed as 'normal', yet where this 'normality' is beset with contradictions and ambiguities. Rather than viewing these as a reflection of resistances to or failures of the enactment of HIV as 'normal', we argue that, insofar as these contradictions are generated by the research interview as a distinct 'talking technology', they emerge as crucial to the normative (re)production of what counts as 'living with HIV' (in the UK) and are an inherent part of the broader performative 'normalisation' of the virus.


Polymeric waveguide Bragg grating filter using soft lithography.
We use the soft lithography technique to fabricate a polymeric waveguide Bragg grating filter. Master grating structure is patterned by e-beam lithography. Using an elastomeric stamp and capillary action, uniform grating structures with very thin residual layers are transferred to the UV curable polymer without the use of an imprint machine. The waveguide layer based on BCB optical polymer is fabricated by conventional optical lithography. This approach provides processing simplicity to fabricate Bragg grating filters.


Prediction Accuracy With Electronic Medical Records Versus Administrative Claims.
The objective of this study was to evaluate the incremental predictive power of electronic medical record (EMR) data, relative to the information available in more easily accessible and standardized insurance claims data. Using both EMR and Claims data, we predicted outcomes for 118,510 patients with 144,966 hospitalizations in 8 hospitals, using widely used prediction models. We use cross-validation to prevent overfitting and tested predictive performance on separate data that were not used for model training. We predict 4 binary outcomes: length of stay (≥7 d), death during the index admission, 30-day readmission, and 1-year mortality. We achieve nearly the same prediction accuracy using both EMR and claims data relative to using claims data alone in predicting 30-day readmissions [area under the receiver operating characteristic curve (AUC): 0.698 vs. 0.711; positive predictive value (PPV) at top 10% of predicted risk: 37.2% vs. 35.7%], and 1-year mortality (AUC: 0.902 vs. 0.912; PPV: 64.6% vs. 57.6%). EMR data, especially from the first 2 days of the index admission, substantially improved prediction of length of stay (AUC: 0.786 vs. 0.837; PPV: 58.9% vs. 55.5%) and inpatient mortality (AUC: 0.897 vs. 0.950; PPV: 24.3% vs. 14.0%). Results were similar for sensitivity, specificity, and negative predictive value across alternative cutoffs and for using alternative types of predictive models. EMR data are useful in predicting short-term outcomes. However, their incremental value for predicting longer-term outcomes is smaller. Therefore, for interventions that are based on long-term predictions, using more broadly available claims data is equally effective.


Disease-specific out-of-pocket healthcare expenditure in urban Bangladesh: A Bayesian analysis.
Because of the rapid increase of non-communicable diseases (NCDs) and high burden of healthcare-related financial issues in Bangladesh, there is a concern that out-of-pocket (OOP) payments related to illnesses may become a major burden on household. It is crucial to understand what are the major illnesses responsible for high OPP at the household level to help policymakers prioritize key areas of actions to protect the household from 100% financial hardship for seeking health care as part of universal health coverage. We first estimated the costs of illnesses among a population in urban Bangladesh, and then assessed the household financial burden associated with these illnesses. A cross-sectional survey of 1593 randomly selected households was carried out in Bangladesh (urban area of Rajshahi city), in 2011. Catastrophic expenditure was estimated at 40% threshold of household capacity to pay. We employed the Bayesian two-stage hurdle model and Bayesian logistic regression model to estimate age-adjusted average cost and the incidence of household financial catastrophe for each illness, respectively. Overall, approximately 45% of the population of Bangladesh had at least one episode of illness. The age-sex-adjusted average medical expenses and catastrophic health care expenditure among the households were TK 621 and 8%, respectively. Households spent the highest amount of money 7676.9 on paralysis followed by liver disease (TK 2695.4), injury (TK 2440.0), mental disease (TK 2258.0), and tumor (TK 2231.2). These diseases were also responsible for higher incidence of financial catastrophe. Our study showed that 24% of individuals who suffered typhoid incurred catastrophic expenditure followed by liver disease (12.3%), tumor (12.1%), heart disease (8.4%), injury (7.9%), mental disease (7.9%), cataract (7.1%), and paralysis (6.5%). The study findings suggest that chronic illnesses were responsible for high costs and high catastrophic expenditures in Bangladesh. Effective risk pooling mechanism might reduce household financial burden related to illnesses. Chronic illness related to NCDs is the major cause of OOP. It is also important to consider prioritizing vulnerable population by subsidizing the high health care cost for some of the chronic illnesses.


Dietary flavonoids and respiratory diseases: a population-based multi-case-control study in Italian adults.
To analyse the associations between chronic respiratory diseases and intakes of total flavonoids and their major subclasses (flavanones, anthocyanins, flavan-3-ols, flavonols, flavones, polymers and proanthocyanidins). Multi-case-control study. The analysis was conducted in the frame of the Genes Environment Interaction in Respiratory Diseases (GEIRD) study. The European Prospective Investigation into Cancer and Nutrition FFQ was used to ascertain dietary intake. Multinomial regression models adjusting for age, sex, centre, BMI, smoking habit, alcohol intake, education, total energy intake, vitamin C intake and total fruit intake were used to examine the associations between dietary exposures and the relative risk ratio (RRR) of being a case. Individuals (n 990) hierarchically defined as follows: cases with asthma (current, n 159; past, n 78), chronic bronchitis (n 47), rhinitis (allergic rhinitis, n 167; non-allergic rhinitis, n 142) and controls (n 97). An increase of 1 sd in flavanones was associated with a reduced risk of non-allergic rhinitis (adjusted RRR = 0·68, 95 % CI 0·47, 0·97); a similar result was found comparing the highest v. lowest quartile of flavanones intake (adjusted RRR = 0·24, 95 % CI 0·10, 0·59). Flavonoids contained in fruits and vegetables, especially flavanones, might reduce the risk of non-allergic rhinitis. No associations were found between other flavonoids and the considered outcomes.


Dissecting the structural basis for the intracellular delivery of OSW-1 by fluorescent probes.
The structural basis for the intracellular delivery of OSW-1 is investigated using fluorescent derivatives of OSW-1 and its closely related congeners. Despite the large differences in activity, all the fluorescent probes are found to translocate across the plasma membrane to the ER and Golgi apparatus. This observation suggests that the glycosylated cholestane moiety plays an important role in the cell internalization and intracellular localization property of OSW-1.


Purification and characterization of thin, aggregative fimbriae from Salmonella enteritidis.
Novel fimbriae were isolated and purified from the human enteropathogen Salmonella enteritidis 27655. These fimbriae were thin (measuring 3 to 4 nm in diameter), were extremely aggregative, and remained cell associated despite attempts to separate them from blended cells by centrifugation. The thin fimbriae were not solubilized in 5 M NaOH or in boiling 0.5% deoxycholate, 8 M urea, or 1 to 2% sodium dodecyl sulfate (SDS) with or without 5% beta-mercaptoethanol. Therefore, an unconventional purification procedure based on the removal of contaminating cell macromolecules in sonicated cell extracts by enzymatic digestion and preparative SDS-polyacrylamide gel electrophoresis (PAGE) was used. The insoluble fimbriae recovered from the well of the gel required depolymerization in formic acid prior to analysis by SDS-PAGE. Acid depolymerization revealed that the fimbriae were composed of fimbrin subunits, each with an apparent molecular mass of 17 kDa. Although their biochemical characteristics and amino acid composition were typical of fimbriae in general, these thin fimbriae were clearly distinct from other previously characterized fimbriae. Moreover, their fimbrin subunits had a unique N-terminal amino acid sequence. Native fimbriae on whole cells were specifically labeled with immune serum raised to the purified fimbriae. This immune serum also reacted with the denatured 17-kDa fimbrin protein in Western blots. The polyclonal immune serum did not cross-react with the other two native fimbrial types produced by this strain or with their respective fimbrins on Western blots (immunoblots). Therefore, these fimbriae represent the third fimbrial type produced by the enteropathogen S. enteritidis.


Finishing a whole-genome shotgun: release 3 of the Drosophila melanogaster euchromatic genome sequence.
The Drosophila melanogaster genome was the first metazoan genome to have been sequenced by the whole-genome shotgun (WGS) method. Two issues relating to this achievement were widely debated in the genomics community: how correct is the sequence with respect to base-pair (bp) accuracy and frequency of assembly errors? And, how difficult is it to bring a WGS sequence to the accepted standard for finished sequence? We are now in a position to answer these questions. Our finishing process was designed to close gaps, improve sequence quality and validate the assembly. Sequence traces derived from the WGS and draft sequencing of individual bacterial artificial chromosomes (BACs) were assembled into BAC-sized segments. These segments were brought to high quality, and then joined to constitute the sequence of each chromosome arm. Overall assembly was verified by comparison to a physical map of fingerprinted BAC clones. In the current version of the 116.9 Mb euchromatic genome, called Release 3, the six euchromatic chromosome arms are represented by 13 scaffolds with a total of 37 sequence gaps. We compared Release 3 to Release 2; in autosomal regions of unique sequence, the error rate of Release 2 was one in 20,000 bp. The WGS strategy can efficiently produce a high-quality sequence of a metazoan genome while generating the reagents required for sequence finishing. However, the initial method of repeat assembly was flawed. The sequence we report here, Release 3, is a reliable resource for molecular genetic experimentation and computational analysis.


A telemetry study shows that an endangered nocturnal avian species roosts in extremely dry habitats to avoid predation.
Describing animal space use is essential for understanding their ecological needs and for planning effective conservation schemes. Notably, certain biomes and life histories are understudied due to methodological challenges in tracking animals in their natural habitats. Specifically, both arid environments and nocturnal species are not sufficiently researched compared to diurnal species and to other biomes. This knowledge gap hinders our ability to properly prioritize habitats for species protection in areas undergoing human-related development. Here, we investigate the movement ecology of the Egyptian Nightjar (Caprimulgus aegyptius) in the arid Dead-sea region of Israel, the Palestinian Authority (the West Bank) and Jordan. This nocturnal insectivore is a cryptic desert-dweller and was considered locally extinct until it was rediscovered in 2016. For this work we tracked twelve individuals using GPS tags to determine how this resource-poor environment affects their home range, (predicting large areas), habitat use, and day-roost ecology. We found that the tracked Egyptian Nightjars had a much larger home range area than other Nightjar species, commuting nightly between foraging grounds and day-roosts. We found, as expected, intensive foraging activity at agricultural fields, where artificial irrigation likely supports higher resource (insect) density. Additionally, we found that individuals showed very high roosting site fidelity, often returning to the same specific site, located in extremely dry and exposed habitats, presumably for predator avoidance. This finding highlights the ecological value of these barren habitats that are often considered "lifeless" and therefore of lower priority for conservation. Consequently, our research demonstrates the importance of describing the space-use of nocturnal animals in arid habitats for conservation efforts.


Excess Dermal Tissue Remodeling In Vivo: Does It Settle?
Surgical manipulation of skin may result in undesired puckering of excess tissue, which is generally assumed to settle over time. In this article, the authors address the novel question of how this excess tissue remodels. Purse-string sutures (6-0 nylon) were placed at the midline dorsum of 22 wild-type BALB/c mice in a circular pattern marked with tattoo ink. Sutures were cinched and tied under tension in the treatment group, creating an excess tissue deformity, whereas control group sutures were tied without tension. After 2 or 4 weeks, sutures were removed. The area of tattooed skin was measured up to 56 days after suture removal. Histologic analysis was performed on samples harvested 14 days after suture removal. The majority of excess tissue deformities flattened within 2 days after suture removal. However, the sutured skin in the treatment group decreased in area by an average of 18 percent from baseline (n = 9), compared to a 1 percent increase in the control group (n = 10) at 14 days after suture removal (p < 0.05). This was similarly observed at 28 days (treatment, -11.7 percent; control, 4.5 percent; n = 5; p = 0.0243). Despite flattening, deformation with purse-string suture correlated with increased collagen content of skin, in addition to increased numbers of myofibroblasts. Change in area did not correlate with duration of suture placement. Excess dermal tissue deformities demonstrate the ability to remodel with gross flattening of the skin, increased collagen deposition, and incomplete reexpansion to baseline area. Further studies will reveal whether our findings in this mouse model translate to humans.


Identifying artemisinin resistance from parasite clearance half-life data with a simple Shiny web application.
The emergence of artemisinin-resistant Plasmodium falciparum malaria is a major threat to malaria elimination. New tools for supporting the surveillance of artemisinin resistance are critical for current and future malaria control and elimination strategies. We have developed an open-access, user-friendly, web-based tool to analyse parasite clearance half-life data of P. falciparum infected patients after treatment with artemisinin derivatives, so that resistance to artemisinin can be identified. The tool can be accessed at bit.ly/id_artemisinin_resistance.


A Single Amino Acid Substitution in the M Protein Attenuates Japanese Encephalitis Virus in Mammalian Hosts.
Japanese encephalitis virus (JEV) membrane (M) protein plays important structural roles in the processes of fusion and maturation of progeny virus during cellular infection. The M protein is anchored in the viral membrane, and its ectodomain is composed of a flexible N-terminal loop and a perimembrane helix. In this study, we performed site-directed mutagenesis on residue 36 of JEV M protein and showed that the resulting mutation had little or no effect on the entry process but greatly affected virus assembly in mammalian cells. Interestingly, this mutant virus had a host-dependent phenotype and could develop a wild-type infection in insect cells. Experiments performed on infectious virus as well as in a virus-like particle (VLP) system indicate that the JEV mutant expresses structural proteins but fails to form infectious particles in mammalian cells. Using a mouse model for JEV pathogenesis, we showed that the mutation conferred complete attenuation in vivo. The production of JEV neutralizing antibodies in challenged mice was indicative of the immunogenicity of the mutant virus in vivo. Together, our results indicate that the introduction of a single mutation in the M protein, while being tolerated in insect cells, strongly impacts JEV infection in mammalian hosts. JEV is a mosquito-transmitted flavivirus and is a medically important pathogen in Asia. The M protein is thought to be important for accommodating the structural rearrangements undergone by the virion during viral assembly and may play additional roles in the JEV infectious cycle. In the present study, we show that a sole mutation in the M protein impairs the JEV infection cycle in mammalian hosts but not in mosquito cells. This finding highlights differences in flavivirus assembly pathways among hosts. Moreover, infection of mice indicated that the mutant was completely attenuated and triggered a strong immune response to JEV, thus providing new insights for further development of JEV vaccines.


Dutch-Flemish translation of 17 item banks from the patient-reported outcomes measurement information system (PROMIS).
The Patient-Reported Outcomes Measurement Information System (PROMIS(®)) is a new, state-of-the-art assessment system for measuring patient-reported health and well-being of adults and children that has the potential to be more valid, reliable and responsive than existing PROMs. The PROMIS items can be administered in short forms or, more efficiently, through computerized adaptive testing. This paper describes the translation of 563 items from 17 PROMIS item banks (domains) for adults from the English source into Dutch-Flemish. The translation was performed by FACITtrans using standardized methodology and approved by the PROMIS Statistical Center. The translation included four forward translations, two back-translations, three to five independent reviews (at least two Dutch, one Flemish) and pre-testing in 70 adults (age range 20-77) from the Netherlands and Flanders. A small number of items required separate translations for Dutch and Flemish: physical function (five items), pain behaviour (two items), pain interference (one item), social isolation (one item) and global health (one item). Challenges faced in the translation process included: scarcity or overabundance of possible translations, unclear item descriptions, constructs broader/smaller in the target language, difficulties in rank ordering items, differences in unit of measurement, irrelevant items or differences in performance of activities. By addressing these challenges, acceptable translations were obtained for all items. The methodology used and experience gained in this study can be used as an example for researchers in other countries interested in translating PROMIS. The Dutch-Flemish PROMIS items are linguistically equivalent. Short forms will soon be available for use and entire item banks are ready for cross-cultural validation in the Netherlands and Flanders.


Reduction of placental nitric oxide synthase activity in pre-eclampsia.
1. The role of the potent vasodilator nitric oxide in the pathogenesis of pre-eclampsia is unclear. We have tested the hypothesis that placental activity of the enzyme which synthesizes nitric oxide (nitric oxide synthase) is reduced in pre-eclampsia. 2. Placentae were obtained after vaginal delivery or Caesarean section from women who had been assigned to the following groups according to standard obstetric criteria: term non-pre-eclamptic control, term pre-eclamptic, preterm non-pre-eclamptic control and preterm pre-eclamptic. Nitric oxide synthase activity of placental tissue homogenates was assessed by measuring conversion of [3H]L-arginine into [3H]L-citrulline in the presence of NADPH, FAD, tetrahydrobiopterin, calmodulin, CaCl2, magnesium acetate and a range of L-arginine concentrations. Michaelis Menton constants (K(m)) amd maximum velocities of reaction (Vmax) were calculated using Lineweaver-Burk analysis. 3. Vmax was significantly reduced in both term and preterm pre-eclamptic placentae compared with placentae from corresponding gestation-matched controls. There were no significant differences in the K(m) values for nitric oxide synthase between any of the four groups, nor were Vmax or K(m) values significantly influenced by mode of delivery. 4. These results provide evidence that human placental nitric oxide synthase activity is significantly reduced in pre-eclampsia. Such a reduction was evident at both term and preterm gestations. Reduced placental nitric oxide synthase activity may have an adverse effect on placental haemodynamic function in pre-eclampsia, and could be involved in the pathogenesis of this important and common obstetric complication.


Transcription factors interfering with dedifferentiation induce cell type-specific transcriptional profiles.
Transcription factors (TFs) are able to regulate differentiation-related processes, including dedifferentiation and direct conversion, through the regulation of cell type-specific transcriptional profiles. However, the functional interactions between the TFs regulating different transcriptional profiles are not well understood. Here, we show that the TFs capable of inducing cell type-specific transcriptional profiles prevent the dedifferentiation induced by TFs for pluripotency. Of the large number of TFs expressed in a neural-lineage cell line, we identified a subset of TFs that, when overexpressed, strongly interfered with the dedifferentiation triggered by the procedure to generate induced pluripotent stem cells. This interference occurred through a maintenance mechanism of the cell type-specific transcriptional profile. Strikingly, the maintenance activity of the interfering TF set was strong enough to induce the cell line-specific transcriptional profile when overexpressed in a heterologous cell type. In addition, the TFs that interfered with dedifferentiation in hepatic-lineage cells involved TFs with known induction activity for hepatic-lineage cells. Our results suggest that dedifferentiation suppresses a cell type-specific transcriptional profile, which is primarily maintained by a small subset of TFs capable of inducing direct conversion. We anticipate that this functional correlation might be applicable in various cell types and might facilitate the identification of TFs with induction activity in efforts to understand differentiation.


Evaluation of curcuma and ginger mixture ability to prevent ROS production induced by bisphenol S: an in vitro study.
The use of bisphenol S (BPS) as a substitute of Bisphenol A is increasing in several products and it can be found in different environmental and biological matrices. Its toxicity has been studied at different levels and one of BPS toxic mechanisms at high concentrations seems to be the induction of oxidative stress through the generation of reactive oxygen species (ROS). This study evaluates the ability of a curcuma and ginger (CG) mixture to exert an antioxidant effect on rat hepatocytes treated with BPS. The effects of the mixture were compared to those of a well-known antioxidant (Trolox). Three different BPS concentrations were used in order to verify ROS production. 70 µg/mL and 150 µg/mL of BPS generated a significant ROS increase (p < 0.01) as compared to control, while CG mixture was able to decrease this ROS production in hepatic cells, as compared to cells treated with 70 µg/ml of BPS (p < 0.01) restoring control levels. BPS 70 µg/mL was tested for total antioxidant capacity (TEAC), superoxide dismutase (SOD) and total thiols. TEAC and SOD significant decreased (p < 0.05 and p < 0.01, respectively) as compared to controls and CG mixture was able to restore control values. Given the widespread BPS use, results obtained in this study can be of high impact for the community, demonstrating the ability of a mixture of natural products to prevent BPS-induced oxidative stress.


Hemorrhagic complications of anticoagulant and thrombolytic treatment: American College of Chest Physicians Evidence-Based Clinical Practice Guidelines (8th Edition).
This article about hemorrhagic complications of anticoagulant and thrombolytic treatment is part of the Antithrombotic and Thrombolytic Therapy: American College of Chest Physicians Evidence-Based Clinical Practice Guidelines (8th Edition). Bleeding is the major complication of anticoagulant and fibrinolytic therapy. The criteria for defining the severity of bleeding vary considerably between studies, accounting in part for the variation in the rates of bleeding reported. The major determinants of vitamin K antagonist (VKA)-induced bleeding are the intensity of the anticoagulant effect, underlying patient characteristics, and the length of therapy. There is good evidence that VKA therapy, targeted international normalized ratio (INR) of 2.5 (range, 2.0-3.0), is associated with a lower risk of bleeding than therapy targeted at an INR > 3.0. The risk of bleeding associated with IV unfractionated heparin (UFH) in patients with acute venous thromboembolism is < 3% in recent trials. This bleeding risk may increase with increasing heparin dosages and age (> 70 years). Low-molecular-weight heparin (LMWH) is associated with less major bleeding compared with UFH in acute venous thromboembolism. Higher doses of UFH and LMWH are associated with important increases in major bleeding in ischemic stroke. In ST-segment elevation myocardial infarction, addition of LMWH, hirudin, or its derivatives to thrombolytic therapy is associated with a small increase in the risk of major bleeding, whereas treatment with fondaparinux or UFH is associated with a lower risk of bleeding. Thrombolytic therapy increases the risk of major bleeding 1.5-fold to threefold in patients with acute venous thromboembolism, ischemic stroke, or ST-elevation myocardial infarction.


Q-methodology in nursing research: a promising method for the study of subjectivity.
This article provides an overview and application of Q-methodology for nursing researchers, with an illustration of its appropriate usage. Q-methodology has been identified as a method for the analysis of subjective viewpoints and has the strengths of both qualitative and quantitative methods. It shares with qualitative methodologies the aim of exploring subjectivity; however, statistical techniques are used to reveal the structure of views. This article describes the use of Q-methodology to examine subjectivity systematically, revealing connections between accounts that other techniques may overlook. An example from the literature is presented. Q-methodology is useful in qualitative nursing research concerned with the exploration and comparison of subjectivity and attitudes. It can be used to effectively identify attitudes, perceptions, feelings, and values as well as explore life experiences such as stress, self-esteem, body image, and satisfaction.


Rapid culture-independent microbial analysis aboard the international space station (ISS) stage two: quantifying three microbial biomarkers.
Abstract A portable, rapid, microbial detection unit, the Lab-On-a-Chip Application Development Portable Test System (LOCAD-PTS), was launched to the International Space Station (ISS) as a technology demonstration unit in December 2006. Results from the first series of experiments designed to detect Gram-negative bacteria on ISS surfaces by quantifying a single microbial biomarker lipopolysaccharide (LPS) were reported in a previous article. Herein, we report additional technology demonstration experiments expanding the on-orbit capabilities of the LOCAD-PTS to detecting three different microbial biomarkers on ISS surfaces. Six different astronauts on more than 20 occasions participated in these experiments, which were designed to test the new beta-glucan (fungal cell wall molecule) and lipoteichoic acid (LTA; Gram-positive bacterial cell wall component) cartridges individually and in tandem with the existing Limulus Amebocyte Lysate (LAL; Gram-negative bacterial LPS detection) cartridges. Additionally, we conducted the sampling side by side with the standard culture-based detection method currently used on the ISS. Therefore, we present data on the distribution of three microbial biomarkers collected from various surfaces in every module present on the ISS at the time of sampling. In accordance with our previous experiments, we determined that spacecraft surfaces known to be frequently in contact with crew members demonstrated higher values of all three microbial molecules. Key Words: Planetary protection-Spaceflight-Microbiology-Biosensor. Astrobiology 12, 830-840.


Two excellent phase-matchable infrared nonlinear optical materials based on 3D diamond-like frameworks: RbGaSn2Se6 and RbInSn2Se6.
Mid- and far-infrared (MFIR) nonlinear optical (NLO) crystals with excellent performances are critical to laser frequency-conversion technology. However, the current commercial MFIR NLO crystals, including AgGaS2 (AGS), AgGaSe2 and ZnGeP2, suffer from certain intrinsic drawbacks and cannot achieve a good balance between large second-harmonic generation (SHG) efficiency and high laser-induced damage thresholds (LIDTs). Herein, we report two new phase-matchable MFIR NLO chalcogenides, specifically RbXSn2Se6 (X = Ga, In), which were successfully synthesized by high-temperature solid-state reactions. The remarkable structural feature of these materials was their 3D diamond-like framework (DLF) stacked by M3Se9 (M = X/Sn) asymmetric building units of vertex-sharing MSe4 tetrahedra along the c axis. Significantly, both of the materials showed the excellent NLO performances with the desired balance between their large SHG efficiencies (4.2 and 4.8 × benchmark AGS) and large LIDTs (8.9 and 8.1 × benchmark AGS), demonstrating that the title compounds meet the crucial conditions as promising MFIR NLO candidates. Furthermore, the crystal structures, synthesis, and theoretical analysis, as well as optical properties are presented herein.


Steering Molecular Activity with Optogenetics: Recent Advances and Perspectives.
Optogenetics utilizes photosensitive proteins to manipulate the localization and interaction of molecules in living cells. Because light can be rapidly switched and conveniently confined to the sub-micrometer scale, optogenetics allows for controlling cellular events with an unprecedented resolution in time and space. The past decade has witnessed an enormous progress in the field of optogenetics within the biological sciences. The ever-increasing amount of optogenetic tools, however, can overwhelm the selection of appropriate optogenetic strategies. Considering that each optogenetic tool may have a distinct mode of action, a comparative analysis of the current optogenetic toolbox can promote the further use of optogenetics, especially by researchers new to this field. This review provides such a compilation that highlights the spatiotemporal accuracy of current optogenetic systems. Recent advances of optogenetics in live cells and animal models are summarized, the emerging work that interlinks optogenetics with other research fields is presented, and exciting clinical and industrial efforts to employ optogenetic strategy toward disease intervention are reported.


Importance of discriminator base stacking interactions: molecular dynamics analysis of A73 microhelix(Ala) variants.
Transfer of alanine from Escherichia coli alanyl-tRNA synthetase (AlaRS) to RNA minihelices that mimic the amino acid acceptor stem of tRNA(Ala) has been shown, by analysis of variant minihelix aminoacylation activities, to involve a transition state sensitive to changes in the 'discriminator' base at position 73. Solution NMR has indicated that this single-stranded nucleotide is predominantly stacked onto G1 of the first base pair of the alanine acceptor stem helix. We report the activity of a new variant with the adenine at position 73 substituted by its non-polar isostere 4-methylindole (M). Despite lacking N7, this analog is well tolerated by AlaRS. Molecular dynamics (MD) simulations show that the M substitution improves position 73 base stacking over G1, as measured by a stacking lifetime analysis. Additional MD simulations of wild-type microhelix(Ala) and six variants reveal a positive correlation between N73 base stacking propensity over G1 and aminoacylation activity. For the two DeltaN7 variants simulated we found that the propensity to stack over G1 was similar to the analogous variants that contain N7 and we conclude that the decrease in aminoacylation efficiency observed upon deletion of N7 is likely due to loss of a direct stabilizing interaction with the synthetase.


Contact burns: the influence of agents and mechanisms of injury on anatomical burn locations in children <5 years old and associations with child protection referrals.
To identify how causative agents and mechanisms of injury influence the location of an accidental contact burn in children and whether these factors differ in cases referred for child protection (CP) assessment. Prospective multicentre cross-sectional study. 20 hospital sites across England and Wales, including: emergency departments, minor injury units and regional burn units. Children less than 5 years old who attended hospital for a contact burn (August 2015 to September 2018). Location of burns with respect to agent and mechanism for accidental contact burns. Secondary outcome: mechanism, agent and location of burns referred for CP assessment. 816 accidental burns and 92 referrals for CP assessment. The most common for accidental burns: mechanism was reaching while stationary (68%, 553/816), agent was oven (24.5%, 200/816) and site was the hand (69.2%, 565/816). Burns to head and trunk were rare at 3.7% (30/816). The data enabled a tabulation of the locations of burns as predicted by agent and mechanism of injury. The location of the burn was most strongly influenced by mechanism.Burns from irons (p<0.01), caused by mechanisms independent of the child (p=0.01), unwitnessed burns (p<0.001) and burns to the head and trunk (p<0.001) were significantly more common among the children referred for CP assessment. By overlaying agent, mechanism and site it was possible to tabulate and quantify simple narratives of accidental contact burns in population of young children. These findings have the potential to aid clinicians in recognising accidental contact burns.


Quantifying intermittent transport in cell cytoplasm.
Active cellular transport is a fundamental mechanism for protein and vesicle delivery, cell cycle, and molecular degradation. Viruses can hijack the transport system and use it to reach the nucleus. Most transport processes consist of intermittent dynamics, where the motion of a particle, such as a virus, alternates between pure Brownian and directed movement along microtubules. In this Rapid Communication, we estimate the mean time for a particle to attach to a microtubule network. This computation leads to a coarse grained equation of the intermittent motion in radial and cylindrical geometries. Finally, by using the degradation activity inside the cytoplasm, we obtain refined asymptotic estimations for the probability and the mean time a virus reaches a small nuclear pore.


Chronic oral treatment with risperidone impairs recognition memory and alters brain-derived neurotrophic factor and related signaling molecules in rats.
Antipsychotic drugs (APDs) are essential for the treatment of schizophrenia and other neuropsychiatric illnesses such as bipolar disease. However, they are also extensively prescribed off-label for many other conditions, a practice that is controversial given their potential for long-term side effects. There is clinical and preclinical evidence that chronic treatment with some APDs may lead to impairments in cognition and decreases in brain volume, although the molecular mechanisms of these effects are unknown. The purpose of the rodent studies described here was to evaluate a commonly prescribed APD, risperidone, for chronic effects on recognition memory, brain-derived neurotrophic factor (BDNF), its precursor proBDNF, as well as relevant downstream signaling molecules that are known to influence neuronal plasticity and cognition. Multiple cohorts of adult rats were treated with risperidone (2.5 mg/kg/day) or vehicle (dilute acetic acid solution) in their drinking water for 30 or 90 days. Subjects were then evaluated for drug effects on recognition memory in a spontaneous novel object recognition task and protein levels of BDNF-related signaling molecules in the hippocampus and prefrontal cortex. The results indicated that depending on the treatment period, a therapeutically relevant daily dose of risperidone impaired recognition memory and increased the proBDNF/BDNF ratio in the hippocampus and prefrontal cortex. Risperidone treatment also led to a decrease in Akt and CREB phosphorylation in the prefrontal cortex. These results indicate that chronic treatment with a commonly prescribed APD, risperidone, has the potential to adversely affect recognition memory and neurotrophin-related signaling molecules that support synaptic plasticity and cognitive function.


Prescribed regimen intensity in diverse youth with type 1 diabetes: role of family and provider perceptions.
Recent literature suggests that disparities in prescribed treatments may exist for youth with type 1 diabetes. There is limited research to date examining factors associated with prescribed regimen intensity in this population. In this study, we examined racial/ethnic differences in regimen intensity and predictors of regimen intensity in youth with type 1 diabetes. We expected that minority youth would have less intensive regimens and that caregiver and physician perceptions would be associated with regimen intensity. This cross-sectional study included 178 families of 10- to 17-yr-old youth at three endocrinology clinics. Caregivers reported perceived costs and benefits of intensive regimens. Physicians described the prescribed treatment and their perceptions of family/child competence and self-management. Analyses included analysis of covariance and hierarchical multiple linear regression. Findings indicate a disparity in regimen intensity for minority youth. Caregiver perceptions of costs associated with intensive regimens and physician perceptions of family competence are associated with prescribed regimen intensity. Interventions targeting disparities in prescribed regimen intensity should be considered. Further research is needed to understand the role of family perceptions of treatments and physician clinical decision making in addressing health disparities in type 1 diabetes.


Characterizing a Foraging Hotspot for Short-Finned Pilot Whales and Blainville's Beaked Whales Located off the West Side of Hawai'i Island by Using Tagging and Oceanographic Data.
Satellite tagging data for short-finned pilot whales (Globicephala macrorhynchus) and Blainville's beaked whales (Mesoplodon densirostris) were used to identify core insular foraging regions off the Kona (west) Coast of Hawai'i Island. Ship-based active acoustic surveys and oceanographic model output were used in generalized additive models (GAMs) and mixed models to characterize the oceanography of these regions and to examine relationships between whale density and the environment. The regions of highest density for pilot whales and Blainville's beaked whales were located between the 1000 and 2500 m isobaths and the 250 and 2000 m isobaths, respectively. Both species were associated with slope waters, but given the topography of the area, the horizontal distribution of beaked whales was narrower and located in shallower waters than that of pilot whales. The key oceanographic parameters characterizing the foraging regions were bathymetry, temperature at depth, and a high density of midwater micronekton scattering at 70 kHz in 400-650 m depths that likely represent the island-associated deep mesopelagic boundary community and serve as prey for the prey of the whales. Thus, our results suggest that off the Kona Coast, and potentially around other main Hawaiian Islands, the deep mesopelagic boundary community is key to a food web that supports insular cetacean populations.


Association between diabetes, severe hypoglycaemia, and electroencephalographic abnormalities.
Serial electroencephalographic recordings were made in 70 diabetic children and findings were related to age at electroencephalography and at diagnosis, duration of diabetes, daily insulin dose, long term metabolic control assessed by glycated haemoglobin A1 (HbA1) concentrations, and severe hypoglycaemic episodes. Abnormalities were found in 18 (26%) of diabetic children, and in only five (7%) of control subjects. There were no associations between electroencephalographic abnormalities and duration of diabetes, daily insulin dose, or HbA1 concentration. Diabetic children with electroencephalographic abnormalities were younger, had an earlier onset of diabetes and 21/34 (62%) of them had previously severe attacks of hypoglycaemia, whereas abnormalities were found in only 13/43 (30%) of diabetic children who had not had severe hypoglycaemia. All diabetic children with hypoglycaemic convulsions had permanent electroencephalographic abnormalities. The degree of metabolic control had no effect on the electroencephalographic findings during the early years of diabetes, but previous severe hypoglycaemia, young age, and early onset seem to be important risk factors for electroencephalographic abnormalities.


Complex formation of yeast Rev1 with DNA polymerase eta.
In Saccharomyces cerevisiae, Rev1 functions in translesion DNA synthesis (TLS) together with polymerase zeta (Pol zeta), comprised of the Rev3 catalytic and Rev7 accessory subunits. Rev1 plays an indispensable structural role in promoting Pol zeta function, and deletion of the Rev1-C terminal region that is involved in physical interactions with Rev3 inactivates Pol zeta function in TLS. In humans, however, Rev1 has been shown to physically interact with the Y-family polymerases Pol eta, Pol iota, and Pol kappa, and the Rev1 C terminus mediates these interactions. Since all the available genetic and biochemical evidence in yeast support the requirement of Rev1 as a structural element for Pol zeta and not for Pol eta, these observations have raised the possibility that in its structural role, Rev1 has diverged between yeast and humans. Here we show that although in yeast a stable Rev1-Pol eta complex can be formed, this complex formation involves the polymerase-associated domain of Rev1 and not the Rev1 C terminus as in humans. We also found that the DNA synthesis activity of Rev1 is enhanced in this complex. We discuss the implications of these and other observations for the possible divergence of Rev1's structural role between yeast and humans.


Hepatitis C virus infection: 'beyond the liver'.
There are rare reports of association between hepatitis C virus (HCV) infection and dermatomyositis although cause and effect remains to be proven. We present a clinical case with a probable cause and effect association between these two entities. A 71-year-old woman developed an erythematous exanthem with pruritic and scaly lesions located at the torso and upper limbs associated with heliotrope and Gottron's papules. At the same time, she notice a significant loss of muscular strength. Skin and muscular biopsies made the diagnosis of dermatomyositis and the patient started with prednisolone (60 mg/day) with poor symptoms control. Paraneoplastic syndrome, HIV, hepatitis B virus and syphilis infections were excluded. HCV serology was positive, with a viral load of 58 159 IU/mL (genotype 1a). Therefore, the patient underwent a 12-week treatment with grazoprevir 100 mg and elbasvir 50 mg achieving a sustained virological response with regression of skin lesions and complete recovery of muscular strength (photodocumented before/after treatment). Additionally it was possible to reduce prednisolone dosage to 5 mg/day.


Predicting nearest agent distances in artificial worlds.
In a number of multi-agent artificial life studies where agents interact over limited distances, the emergence and/or evolution of a specific behavior may depend critically upon interagent distances. Little theoretical analysis has been done previously concerning how to predict such distances. In this paper, we derive a probabilistic method that, for an agent at an arbitrary location in a two-dimensional cellular world, predicts the expected distance to a nearest other agent. Our method works for many world topologies, and we apply it to determine the expected distance for six commonly used ones. Further, the method is readily adapted to handle special restrictions. Over a wide variety of agent densities we show that the theoretically predicted distances are largely in agreement with the distances measured in computational experiments with randomly placed agents. We then utilize our prediction method to interpret recent observations that an imprecise threshold in the density of agents exists for the evolution of communication. We thus illustrate that, despite its conceptual simplicity, our method can aid the analysis and even the design of complex artificial environments populated by agents that have the potential to interact with one another.


Wireless Monitoring of Changes in Crew Relations during Long-Duration Mission Simulation.
Group structure and cohesion along with their changes over time play an important role in the success of missions where crew members spend prolonged periods of time under conditions of isolation and confinement. Therefore, an objective system for unobtrusive monitoring of crew cohesion and possible individual stress reactions is of high interest. For this purpose, an experimental wireless group structure (WLGS) monitoring system integrated into a mobile psychophysiological system was developed. In the presented study the WLGS module was evaluated separately in six male subjects (27-38 years old) participating in a 520-day simulated mission to Mars. Two days per week, each crew member wore a small sensor that registered the presence and distance of the sensors either worn by the other subjects or strategically placed throughout the isolation facility. The registration between two sensors was on average 91.0% in accordance. A correspondence of 95.7% with the survey video on day 475 confirmed external reliability. An integrated score of the "crew relation time index" was calculated and analyzed over time. Correlation analyses of a sociometric questionnaire (r = .35-.55, p< .05) and an ethological group approach (r = .45-.66, p < 05) provided initial evidence of the method's validity as a measure of cohesion when taking behavioral and activity patterns into account (e.g. only including activity phases in the afternoon). This confirms our assumption that the registered amount of time spent together during free time is associated with the intensity of personal relationships.


Tetrachloroethene-dehalogenating bacteria.
Tetrachloroethene is a frequent groundwater contaminant often persisting in the subsurface environments. It is recalcitrant under aerobic conditions because it is in a highly oxidized state and is not readily susceptible to oxidation. Nevertheless, at least 15 organisms from different metabolic groups, viz. halorespirators (9), acetogens (2), methanogens (3) and facultative anaerobes (2), that are able to metabolize tetrachloroethene have been isolated as axenic cultures to-date. Some of these organisms couple dehalo-genation to energy conservation and utilize tetrachloroethene as the only source of energy while others dehalogenate tetrachloroethene fortuitously. Halorespiring organisms (halorespirators) utilize halogenated organic compounds as electron acceptors in an anaerobic respiratory process. Different organisms exhibit differences in the final products of tetrachloroethene dehalogenation, some strains convert tetrachloroethene to trichloroethene only, while others also carry out consecutive dehalogenation to dichloroethenes and vinyl chloride. Thus far, only a single organism, 'Dehalococcoides ethenogenes' strain 195, has been isolated which dechlorinates tetrachloroethene all the way down to ethylene. The majority of tetrachloroethene-dehalogenating organisms have been isolated only in the past few years and several of them, i.e., Dehalobacter restrictus, Desulfitobacterium dehalogenans, 'Dehalococcoides ethenogenes', 'Dehalospirillum multivorans', Desulfuromonas chloroethenica, and Desulfomonile tiedjei, are representatives of new taxonomic groups. This contribution summarizes the available information regarding the axenic cultures of the tetrachloroethene-dehalogenating bacteria. The present knowledge about the isolation of these organisms, their physiological characteristics, morphology, taxonomy and their ability to dechlorinate tetrachloroethene is presented to facilitate a comprehensive comparison.


Ontogenetic shifts in plant-plant interactions in a rare cycad within angiosperm communities.
Gymnosperms and angiosperms can co-occur within the same habitats but key plant traits are thought to give angiosperms an evolutionary competitive advantage in many ecological settings. We studied ontogenetic changes in competitive and facilitative interactions between a rare gymnosperm (Dioon sonorense, our target species) and different plant and abiotic neighbours (conspecific-cycads, heterospecific-angiosperms, or abiotic-rocks) from 2007 to 2010 in an arid environment of northwestern Mexico. We monitored survival and growth of seedlings, juveniles, and adults of the cycad Dioon sonorense to evaluate how cycad survival and relative height growth rate (RHGR) responded to intra- and interspecific competition, canopy openness, and nearest neighbour. We tested spatial associations among D. sonorense life stages and angiosperm species and measured ontogenetic shifts in cycad shade tolerance. Canopy openness decreased cycad survival while intraspecific competition decreased survival and RHGR during early ontogeny. Seedling survival was higher in association with rocks and heterospecific neighbours where intraspecific competition was lower. Shade tolerance decreased with cycad ontogeny reflecting the spatial association of advanced stages with more open canopies. Interspecific facilitation during early ontogeny of our target species may promote its persistence in spite of increasing interspecific competition in later stages. We provide empirical support to the long-standing assumption that marginal rocky habitats serve as refugia from angiosperm competition for slow-growing gymnosperms such as cycads. The lack of knowledge of plant-plant interactions in rare or endangered species may hinder developing efficient conservation strategies (e.g. managing for sustained canopy cover), especially under the ongoing land use and climatic changes.


No effects of psychosocial stress on intertemporal choice.
Intertemporal choices - involving decisions which trade off instant and delayed outcomes - are often made under stress. It remains unknown, however, whether and how stress affects intertemporal choice. We subjected 142 healthy male subjects to a laboratory stress or control protocol, and asked them to make a series of intertemporal choices either directly after stress, or 20 minutes later (resulting in four experimental groups). Based on theory and evidence from behavioral economics and cellular neuroscience, we predicted a bidirectional effect of stress on intertemporal choice, with increases in impatience or present bias immediately after stress, but decreases in present bias or impatience when subjects are tested 20 minutes later. However, our results show no effects of stress on intertemporal choice at either time point, and individual differences in stress reactivity (changes in stress hormone levels over time) are not related to individual differences in intertemporal choice. Together, we did not find support for the hypothesis that psychosocial laboratory stressors affect intertemporal choice.


DNA microarray analysis of gene expression in endothelial cells in response to 24-h shear stress.
The recently developed DNA microarray technology provides a powerful and efficient tool to rapidly compare the differential expression of a large number of genes. Using the DNA microarray approach, we investigated gene expression profiles in cultured human aortic endothelial cells (HAECs) in response to 24 h of laminar shear stress at 12 dyn/cm(2). This relatively long-term shearing of cultured HAECs led to the modulation of the expression of a number of genes. Several genes related to inflammation and EC proliferation were downregulated, suggesting that 24-h shearing may keep ECs in a relatively noninflammatory and nonproliferative state compared with static cells. Some genes were significantly upregulated by the 24-h shear stress; these includes genes involved in EC survival and angiogenesis (Tie2 and Flk-1) and vascular remodeling (matrix metalloproteinase 1). These results provide information on the profile of gene expression in shear-adapted ECs, which is the case for the native ECs in the straight part of the aorta in vivo.


Application of participation in clinical practice: Key issues.
Rehabilitation services are increasingly targeting involvement in daily life. In the International Classification of Functioning, Disability and Health this is referred to as "participation". How-ever, questions have arisen regarding the conceptualization of participation, and consensus is lacking. The first phase of this study is a critical review of the literature to detect recurring conceptual problems in the application of participation and how researchers deal with these. The second phase is a systematic review to identify how participation measures are operationalized. The critical review found possible solutions to 4 recurring key limitations: (i) how to deal with ambiguity and vagueness regarding the term "participation"; (ii) how to differentiate between activity and participation; (iii) what is the current empirical knowledge about the subjective aspects of participation; (iv) what are the different ways to measure participation. The systematic review found 18 instruments operationalizing participation in different ways: (i) unidimensional: frequency of performing activities; (ii) unidimensional: limitations in experiencing participation when performing activities; (iii) multidimensional: multiple subjective dimensions when performing activities; and (iv) multidimensional: objective and subjective dimensions. Notwithstanding an increasing body of knowledge, some issues remain unclear and how participation is measured is subject to debate. This results in difficulties in the use of participation in clinical practice. However, insight into the current body of knowledge and awareness of shortcomings might help clinicians who aim to apply participation in practice.


Cdc42 localization and cell polarity depend on membrane traffic.
Cell polarity is essential for cell division, cell differentiation, and most differentiated cell functions including cell migration. The small G protein Cdc42 controls cell polarity in a wide variety of cellular contexts. Although restricted localization of active Cdc42 seems to be important for its distinct functions, mechanisms responsible for the concentration of active Cdc42 at precise cortical sites are not fully understood. In this study, we show that during directed cell migration, Cdc42 accumulation at the cell leading edge relies on membrane traffic. Cdc42 and its exchange factor βPIX localize to intracytosplasmic vesicles. Inhibition of Arf6-dependent membrane trafficking alters the dynamics of Cdc42-positive vesicles and abolishes the polarized recruitment of Cdc42 and βPIX to the leading edge. Furthermore, we show that Arf6-dependent membrane dynamics is also required for polarized recruitment of Rac and the Par6-aPKC polarity complex and for cell polarization. Our results demonstrate influence of membrane dynamics on the localization and activation of Cdc42 and consequently on directed cell migration.


RNA-Seq-Based Comparative Transcriptomics: RNA Preparation and Bioinformatics.
The major transcriptome analysis is the determination of differentially expressed genes across experimental conditions. For this, the next-generation sequencing of RNA (RNA-seq) is an increasingly cost-effective technology for the analysis of transcriptomes with several advantages over gene expression microarrays, such as its higher sensitivity and accuracy, broader dynamic range, and the ability to detect novel transcripts, including noncoding RNA molecules, at nucleotide-level resolution. Although these advantages, many microbiology laboratories have not yet applied RNA-seq analyses to their investigations. The high cost of the equipment for next-generation sequencing is no longer an issue, since this intermediate part of the analysis can be provided by commercial or central services. Here, we detail a protocol for the first part of the analysis, the RNA extraction, and an introductory protocol to the bioinformatics analysis of the sequencing data that generates the differential expression results.


The interplay between online and offline explorations of identity, relationships, and sex: a mixed-methods study with LGBT youth.
Although the Internet is commonly used by lesbian, gay, bisexual, and transgender (LGBT) youth to explore aspects of sexual health, little is known about how this usage relates to offline explorations and experiences. This study used a mixed-methods approach to investigate the interplay between online and offline explorations of multiple dimensions of sexual health, which include sexually transmitted infections, sexual identities, romantic relationships, and sexual behaviors. A diverse community sample of 32 LGBT youth (ages 16-24) completed semi-structured interviews, which were transcribed and then qualitatively coded to identify themes. Results indicated that, although many participants evaluated online sexual health resources with caution, they frequently used the Internet to compensate for perceived limitations in offline resources and relationships. Some participants turned to the Internet to find friends and romantic partners, citing the relative difficulty of establishing offline contact with LGBT peers. Further, participants perceived the Internet as an efficient way to discover offline LGBT events and services relevant to sexual health. These results suggest that LGBT youth are motivated to fill gaps in their offline sexual health resources (e.g., books and personal communications) with online information. The Internet is a setting that can be harnessed to provide support for the successful development of sexual health.


Progesterone and estradiol enhance lipid mediated transfection of Sk-Br-3 mammalian cancer cells.
Cyclodextrin encapsulated beta-estradiol and progesterone were used for enhancement of gene delivery using the breast cancer cell line Sk-Br-3. A non-toxic concentration of cyclodextrin encapsulated sex steroids (50 microM) added to lipid or liposomal transfection led to a 12-fold increase of reporter gene expression (luciferase) with progesterone and an 8-fold increase with estradiol using Lipofectamine Plus mediated transfection. Using the lipid formulation Fugene-6 the results were a 5.5-fold and a 4.5-fold increase respectively. This enhancement could only be observed if the sex steroids were added to the cells before application of the DNA-Fugene complex supporting the evidence that intracellular processes are responsible for the activity of the steroids. The strong differences between progesterone and estradiol in modifying Lipofectamine Plus transfection in Sk-Br-3 cells may to be explained by differences in the distribution of these receptors in the cellular compartments. These results seem to add evidence on the possibility of using sex steroids to increase the efficiency of non-viral vectors for transfection, and may ultimately prove to be relevant to gene therapy in the treatment of breast cancer as well as other solid tumors.